CN103931499A - Tissue culture rapid propagation method for callistemon rigidus - Google Patents
Tissue culture rapid propagation method for callistemon rigidus Download PDFInfo
- Publication number
- CN103931499A CN103931499A CN201410152533.1A CN201410152533A CN103931499A CN 103931499 A CN103931499 A CN 103931499A CN 201410152533 A CN201410152533 A CN 201410152533A CN 103931499 A CN103931499 A CN 103931499A
- Authority
- CN
- China
- Prior art keywords
- medium
- sprouting
- culture
- tissue culture
- cultivation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention discloses a tissue culture rapid propagation method for callistemon rigidus and belongs to the technical field of plant tissue culture. The method comprises the following culture steps: (1) selecting and sterilizing an explant; (2) performing primary culture; (3) performing subculture; (4) performing rooting culture; and (5) performing acclimatization and transplant. According to the culture method, the seedling adaptability is high after acclimatization and transplant, so that the acclimatization and transplant survival rate of tissue culture seedlings of callistemon viminalis is over 95 percent. Compared with conventional seed propagation cutting propagation, the tissue culture rapid propagation method has the advantages that the seedling growing period of the callistemon viminalis is greatly shortened, the propagation efficiency can be greatly improved, the seedling growing period is shortened from 1 year to 3-4 months, the seedling growing batch number per year is improved from 1 to 4-8, and the method has wide market application prospects.
Description
Technical field
The invention belongs to field of plant tissue culture technique, be specifically related to callistemon rigidus quick breeding method for tissue culture.
Background technology
Callistemon rigidus (Callistemon viminalis) originates in Australia, is that Myrtaceae (Myrtaceae) callistemon rigidus belongs to (Callistemon) evergreen shrubs or dungarunga, and its flower image test-tube brush, therefore weighing bottle is brushed tree, red bottle brush again.This plant adaptability is wider, can in warm hot weather, grow, and also can adapt to hotter or colder weather.Be suitable for cultivation in moistening, loose rich soil, but also can be in the soil of barren arid well-grown.The rudiment power of this plant is strong, poor growth, and resistance to pruning, anti-atmospheric pollution power is strong.
As a kind of flowers and trees that have ornamental value, callistemon rigidus strain shape grace, sprig is elongated and curved hangs down, tree crown weeping willow shape, and blade has transparent oil droplet point, that rubs has an aromatic oil smell, spend a dense top axil of being born in, the florescence 3-5 month and October, spike is unique beautiful, safflower layer head time in full bloom, very graceful.For afforestation such as parks, scenic area, residential area, road, can arrange in pairs or groups with various plants, plant in isolated planting, group planting, the sheet mode such as plant, build abundant garden landscape.This plant or good economic tree, have good medical value.
Seminal propagation, cottage propagation and propagation by grafiting are the Sterile culture methods that trees are conventional.Up to the present, the most frequently used propagation method of callistemon rigidus platymiscium is seminal propagation.But on the one hand, once the fruit maturation of weeping branch callistemon rigidus will ftracture immediately, seed is very easily scattered, be difficult to gather, and its seed is superfine little, latter stage of ripening, is longer, and germination rate is lower; On the other hand, the cottage propagation survival rate of weeping branch callistemon rigidus is low; As for propagation by grafiting, need to first cultivate rootstock seedling by seminal propagation or cottage propagation equally.Therefore,, for the good Landscape Trees of this characteristic of weeping branch callistemon rigidus, several conventional Sterile culture methods are all unsuitable for scale application.For this problem, in order to adapt to market demand, be badly in need of setting up a kind of new propagation technique system.
Tissue-culturing quick-propagation is the Fast-propagation nursery stock new method of cultivating based on tissue.At present, little about the research of callistemon rigidus platymiscium tissue-culturing quick-propagation aspect both at home and abroad, comprehensive and systematic researches, even if Australia of country of origin does not set up the tissue culture rapid propagation technique system of any callistemon rigidus platymiscium yet always.Therefore; we study taking the weeping branch callistemon rigidus of this genus as object; to exploring its tissue culture rapid propagation technique system of setting up; for its nursery stock scale, standardization, factorial seedling growth provide technical support; meet need of production, for callistemon rigidus platymiscium, application has great theory and practice meaning.
Summary of the invention
The object of this invention is to provide a kind of callistemon rigidus quick breeding method for tissue culture; grow seedlings technical support is provided for callistemon rigidus scale, standardization, batch production, the whole yearization; for further genetic breeding provides theory and technical foundation, there is important theory and practice meaning for this plant introducing a fine variety with utilization and extention of China.
Order of the present invention is achieved through the following technical solutions.
1. a quick breeding method for tissue culture for callistemon rigidus, comprises the following steps:
1) selection of explant and sterilizing: taking the weeping branch callistemon rigidus branch of raw semi-lignified then as explant, branch is cut into the stem segment with axillary bud of 2～3cm, cuts off blade, clean dust with flowing water, soak 3min with liquid detergent solution, then rinse 2h with flowing water; On superclean bench, first by 70% alcohol-pickled 30s for stem section, shake 25s, removes alcohol and with aseptic water washing 5 times, soak 8min with 0.1% mercuric chloride again, shake once every 2min, and with aseptic water washing 5～7 times, put it in sterile water, soak for subsequent use;
2) first culture: 1～2mm is cut away in explant wound after sterilizing, end is inserted in Y1 medium downward vertically, be transferred in new Y1 medium after cultivation 3～7d, and continuation cultivation 10～15d grows sprouting;
3) sprouting extends and cultivates: in Y2 medium, carry out sprouting and extend and cultivate, be elongated to >=3.0cm of sprouting after 20d being transferred to after the sprouting stem segment base portion blackout Partial Resection growing;
4) shoot proliferation is cultivated: the sprouting after elongation growth is cut into the segment of about 1cm, is proceeded in Y2 medium and carry out shoot proliferation cultivation, breed more than sprouting sprouts and be elongated to 2cm after 25～30d; Shoot proliferation is cultivated algebraically and was no more than for 4 generations;
5) culture of rootage: the sprouting that step 4) is grown to more than 2cm is cut, removes the blade of close medium base portion and is transferred to Y3 medium, after cultivation 15d, grows successively new root;
6) acclimatization and transplants: the tissue culture bottle bottle cap of the plant taking root is opened to 3d; The taking-up seedling of taking root, cleans root medium, running water cultivation 5d, and cover water tumbler with polyethylene plastic bag, keep humidity; The taking-up seedling of taking root, transplants to being equipped with in the basin of matrix, keeps 22～25 DEG C of temperature, and humidity 85～95%, suitably shades,
Wherein, the component of described each medium is:
Y1 medium: MS minimal medium (each composition consumption refers to MS minimal medium explanation below), 30g/L sucrose, 7.5g/L agar powder, 0.1mg/L6-benzyl aminoadenine (6-BA), 0.05mg/L methyl α-naphthyl acetate (NAA), pH is 5.8;
Y2 medium: MS minimal medium, 30g/L sucrose, 7.5g/L agar powder, 1.0mg/L6-BA, 0.2mg/L NAA, pH is 5.8;
Y3 medium: MS minimal medium, 20g/L sucrose, 7.5g/L agar powder, 0.25mg/L indolebutyric acid (IBA), 0.25mg/L NAA that macroelement reduces by half, pH is 5.8;
Wherein, the condition of culture described in step 2-5 is: temperature 23-27 DEG C, irradiate 16h, then camera bellows preservation 8 hour by 1400Lx light intensity every day.
The invention has the beneficial effects as follows: for the very low problem of callistemon rigidus Sterile culture efficiency, set up first the tissue culture rapid propagation technique system of these seeds, make its seedling propagation not be subject to the restriction in time, season, weather and place, greatly shorten growing-seedling period, greatly improve breeding efficiency, growing-seedling period shortened to 3 to 4 months by 1 year, and the lot number of growing seedlings is every year brought up to 4 to 8 batches by 1 batch, for its large scale application provides an effective way.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
Embodiment 1:
A quick breeding method for tissue culture for callistemon rigidus, specific as follows:
1) selection of explant and sterilizing: taking the weeping branch callistemon rigidus branch of raw semi-lignified then as explant, branch is cut into the stem section of 2～3cm, cuts off blade, clean dust with flowing water, soak 3min with liquid detergent solution, then rinse 2h with flowing water; On superclean bench, first by 70% alcohol-pickled 30s for stem section, shake 25s, removes alcohol and with aseptic water washing 5 times, then with 0.1% mercuric chloride immersion 8min, shakes once every 2min, and with aseptic water washing 6 times, put it in sterile water, soak for subsequent use;
2) first culture: 1～2mm is cut away in explant wound after sterilizing, end is inserted in Y1 medium downward vertically, be transferred in new Y1 medium after cultivation 5d, and continuation cultivation 12d grows sprouting;
3) sprouting extends and cultivates: in Y2 medium, carry out sprouting and extend and cultivate, be elongated to >=3.0cm of sprouting after 20d being transferred to after the sprouting stem segment base portion blackout Partial Resection growing;
4) shoot proliferation is cultivated: the sprouting after elongation growth is cut into the segment of about 1cm, is proceeded in Y2 medium and carry out shoot proliferation cultivation, breed more than sprouting sprouts and be elongated to 2cm after 28d; It was 4 generations that shoot proliferation is cultivated algebraically;
5) culture of rootage: the sprouting that step 4) is grown to more than 2cm is cut, removes the blade of close medium base portion and is transferred to Y3 medium, after cultivation 15d, grows successively new root;
6) acclimatization and transplants: the tissue culture bottle bottle cap of the callistemon rigidus plant taking root is opened to 3d; The taking-up seedling of taking root, cleans root medium, running water cultivation 5d, and cover water tumbler with polyethylene plastic bag, keep humidity; The taking-up seedling of taking root, transplants to being equipped with in the basin of matrix, keeps 22～25 DEG C of temperature, and humidity 85～95%, suitably shades, and survival rate reaches 95%.
Wherein, the condition of culture described in step 2-5 is: 25 ± 2 DEG C of temperature, irradiate 16h, then camera bellows preservation 8 hour by 1400Lx light intensity every day.
Wherein, the component of described each medium is:
Y1 medium: MS minimal medium, 30g/L sucrose, 7.5g/L agar powder, 0.1mg/L6-benzyl aminoadenine, 0.05mg/L methyl α-naphthyl acetate, pH is 5.8;
Y2 medium: MS minimal medium, 30g/L sucrose, 7.5g/L agar powder, 1.0mg/L6-benzyl aminoadenine, 0.2mg/L methyl α-naphthyl acetate, pH is 5.8;
Y3 medium: MS minimal medium, 20g/L sucrose, 7.5g/L agar powder, 0.25mg/L indolebutyric acid, 0.25mg/L methyl α-naphthyl acetate that macroelement reduces by half, pH is 5.8;
Content (g/L or mg/L) wherein all refers to that each component accounts for the w/v of total medium, as the 30g/L sucrose in Y1 medium, refers in every 1L Y1 medium containing sucrose 30g.
Described MS minimal medium contains macroelement, trace element, molysite, the large class of organic principle four, and in Y1, Y2 medium, the content that each component of MS minimal medium accounts for total medium is: macroelement is: KNO
3(1900mg/L), NH
4nO
3(1650mg/L), MgSO
47H
2o(370mg/L), KH
2pO
4(170mg/L), CaCl
2(440mg/L); Trace element is: KI(0.83mg/L), H
3bO
3(6.2mg/L), MnSO
4h
2o(16.9mg/L), ZnSO
44H
2o(8.6mg/L), CuSO
45H
2o(0.025mg/L), CoCl
26H
2o(0.025mg/L), Na
2moO
42H
2o(0.25mg/L); Molysite is: FeSO
47H
2o(27.85mg/L), Na
2eDTA(37.25mg/L); Organic principle is: glycine (2.0mg/L), nicotinic acid B3(0.5mg/L), thiamine hydrochloride B1(0.1mg/L), puridoxine hydrochloride B6(0.5mg/L) and, inositol (100mg/L).
Macroelement in Y3 medium reduces by half, i.e. KNO
3, NH
4nO
3, MgSO
47H
2o, KH
2pO
4, CaCl
2content reduce by half, the content of other composition is constant.
Claims (1)
1. a quick breeding method for tissue culture for callistemon rigidus, is characterized in that comprising the following steps:
1) selection of explant and sterilizing: taking the weeping branch callistemon rigidus branch of raw semi-lignified then as explant, branch is cut into the stem segment with axillary bud of 2～3cm, cuts off blade, clean dust with flowing water, soak 3min with liquid detergent solution, then rinse 2h with flowing water; On superclean bench, first by 70% alcohol-pickled 30s for stem section, shake 25s, removes alcohol and with aseptic water washing 5 times, soak 8min with 0.1% mercuric chloride again, shake once every 2min, and with aseptic water washing 5～7 times, put it in sterile water, soak for subsequent use;
2) first culture: 1～2mm is cut away in explant wound after sterilizing, end is inserted in Y1 medium downward vertically, be transferred in new Y1 medium after cultivation 3～7d, and continuation cultivation 10～15d grows sprouting;
3) sprouting extends and cultivates: in Y2 medium, carry out sprouting and extend and cultivate, be elongated to >=3.0cm of sprouting after 20d being transferred to after the sprouting stem segment base portion blackout Partial Resection growing;
4) shoot proliferation is cultivated: the sprouting after elongation growth is cut into the segment of about 1cm, is proceeded in Y2 medium and carry out shoot proliferation cultivation, breed more than sprouting sprouts and be elongated to 2cm after 25～30d; Shoot proliferation is cultivated algebraically and was no more than for 4 generations;
5) culture of rootage: the sprouting that step 4) is grown to more than 2cm is cut, removes the blade of close medium base portion and is transferred to Y3 medium, after cultivation 15d, grows successively new root;
6) acclimatization and transplants: the tissue culture bottle bottle cap of the plant taking root is opened to 3d; The taking-up seedling of taking root, cleans root medium, running water cultivation 5d, and cover water tumbler with polyethylene plastic bag, keep humidity; The taking-up seedling of taking root, transplants to being equipped with in the basin of matrix, keeps 22～25 DEG C of temperature, and humidity 85～95%, suitably shades,
Wherein, the component of described each medium is:
Y1 medium: MS minimal medium, 30g/L sucrose, 7.5g/L agar powder, 0.1mg/L6-benzyl aminoadenine, 0.05mg/L methyl α-naphthyl acetate, pH is 5.8;
Y2 medium: MS minimal medium, 30g/L sucrose, 7.5g/L agar powder, 1.0mg/L6-benzyl aminoadenine, 0.2mg/L methyl α-naphthyl acetate, pH is 5.8;
Y3 medium: MS minimal medium, 20g/L sucrose, 7.5g/L agar powder, 0.25mg/L indolebutyric acid, 0.25mg/L methyl α-naphthyl acetate that macroelement reduces by half, pH is 5.8;
Wherein, the condition of culture described in step 2-5 is: temperature 23-27 DEG C, irradiate 16h, then camera bellows preservation 8 hour by 1400Lx light intensity every day.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410152533.1A CN103931499B (en) | 2014-04-16 | 2014-04-16 | A kind of quick breeding method for tissue culture of callistemon rigidus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410152533.1A CN103931499B (en) | 2014-04-16 | 2014-04-16 | A kind of quick breeding method for tissue culture of callistemon rigidus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103931499A true CN103931499A (en) | 2014-07-23 |
| CN103931499B CN103931499B (en) | 2016-07-06 |
Family
ID=51179590
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410152533.1A Expired - Fee Related CN103931499B (en) | 2014-04-16 | 2014-04-16 | A kind of quick breeding method for tissue culture of callistemon rigidus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103931499B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105145176A (en) * | 2015-09-01 | 2015-12-16 | 张莘蔓 | Method for water culture cutting propagation of callistemon cirtinus |
| CN105724177A (en) * | 2016-05-14 | 2016-07-06 | 崔子扬 | Cultivating method of callistemon viminalis |
| CN106386343A (en) * | 2016-08-31 | 2017-02-15 | 广西棕海园林工程有限公司 | Seedling growing method for Callistemon rigidus |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577942A (en) * | 2011-01-06 | 2012-07-18 | 厦门涌泉集团有限公司 | Tissue culture method of callistemon hybridus 'golden ball' |
| US20120331604P1 (en) * | 2011-05-27 | 2012-12-27 | Graham Brown | Callistemon viminalis plant named 'CV01' |
-
2014
- 2014-04-16 CN CN201410152533.1A patent/CN103931499B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577942A (en) * | 2011-01-06 | 2012-07-18 | 厦门涌泉集团有限公司 | Tissue culture method of callistemon hybridus 'golden ball' |
| US20120331604P1 (en) * | 2011-05-27 | 2012-12-27 | Graham Brown | Callistemon viminalis plant named 'CV01' |
Non-Patent Citations (2)
| Title |
|---|
| 舒婷: "美花红千层的组织培养研究", 《安徽林业科技》 * |
| 龚伟等: "红千层的组织培养与快速繁殖", 《四川大学学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105145176A (en) * | 2015-09-01 | 2015-12-16 | 张莘蔓 | Method for water culture cutting propagation of callistemon cirtinus |
| CN105724177A (en) * | 2016-05-14 | 2016-07-06 | 崔子扬 | Cultivating method of callistemon viminalis |
| CN106386343A (en) * | 2016-08-31 | 2017-02-15 | 广西棕海园林工程有限公司 | Seedling growing method for Callistemon rigidus |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103931499B (en) | 2016-07-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103190347B (en) | Teapot dates tissue culturing method | |
| CN101946703B (en) | Method for regenerating plants of Chinese rose by using leaves as explants | |
| CN103348920B (en) | Rapid propagation method for high quality seedlings of Kyara | |
| CN104521715A (en) | Sealing water planting cutting propagation method for gardenias with flower buds | |
| CN104885773A (en) | Method for rapidly cultivating early shaping tissue culture commodity seedlings of blueberries | |
| CN104542281B (en) | Mediterranean Jia Opulus tissue culture propagation method | |
| CN109287486B (en) | Paphiopedilum seed germination rate improving method and paphiopedilum cultivation method | |
| CN104585037A (en) | Tissue culture rapid-propagation method of beaucarnea recurvata | |
| CN109349105A (en) | A kind of iris tissue-cultured seedling mating system | |
| CN109287487B (en) | Seed germination rate improving method and cultivation method for paphiopedilum makino | |
| CN105532459B (en) | A kind of tissue culture and rapid propagation method of Acer palmatum orange dream | |
| CN104642141A (en) | Gerbera adventitious bud induction and plant regeneration method by using root as explant | |
| CN107079817A (en) | Pocket Lanzhou and Xinjiang kind " excellent pocket is blue " tissue culture and rapid propagation method | |
| CN103583360A (en) | Method for improving Abelia seedling salt tolerance by oriented induction | |
| CN101946704B (en) | Method for regenerating Chinese rose plant by using immature seed as explant | |
| CN103931499B (en) | A kind of quick breeding method for tissue culture of callistemon rigidus | |
| CN108770680A (en) | A kind of method of the sapling multiplication and cultivation of red rosy clouds pocket orchid | |
| CN110651713B (en) | Tissue culture method of clematis' Fuji blue | |
| CN104488722A (en) | Quick propagation method for tissue culture of staurogyne sp | |
| CN104488721B (en) | A kind of quick breeding method for tissue culture of snowflake grass | |
| CN108391591B (en) | Tissue culture and rapid propagation method for tabebuia flavedo | |
| CN102884980B (en) | Quick tissue propagation method of paphiopedilum bellatulum | |
| CN100391333C (en) | Aseptic seedling tissue culturing and test tube seedling hardening off and transplating technology for anthurium andraeanum | |
| CN103858763A (en) | Sinningia speciosa tissue culture propagation and tuber storage and re-propagation method | |
| CN102119664B (en) | Method for culturing muscari botryoides mill with plant tissues |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160706 Termination date: 20170416 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |